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Preparation method and application of norcantharidin-loaded exosome

A technology of norcantharidin and exosomes, applied in the field of loaded norcantharidin exosomes and its preparation, can solve the problems of potential toxicity to be verified, adverse reactions, reduced targeting efficiency, etc., and achieve enhanced drug loading , Improve drug efficacy, improve the effect of encapsulation rate

Active Publication Date: 2021-05-11
GUANGDONG PHARMA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, after a large number of animal and clinical trials, it has been found that nanocarriers also have many disadvantages: as exogenous substances, nanocarriers may be taken up by the immune system of non-target organs before they penetrate the cell membrane, thereby reducing their target organ immunity. Most nanocarriers are not completely biodegradable materials, they have certain cytotoxicity and immunogenicity, and the potential toxicity of their residues remains to be verified[1]
Currently, there are two dosage forms of tablet and injection for clinical use, but due to its rapid metabolism in the human body, it is necessary to frequently increase the number of administrations to maintain the drug concentration in order to achieve the therapeutic effect, and long-term application of large doses has adverse effects on the hematopoietic system, urinary system, etc. System, etc. are prone to serious adverse reactions

Method used

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  • Preparation method and application of norcantharidin-loaded exosome
  • Preparation method and application of norcantharidin-loaded exosome
  • Preparation method and application of norcantharidin-loaded exosome

Examples

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Embodiment 1

[0067] Example 1: A method for preparing norcantharidin-loaded exosomes and its characterization

[0068] (1) Extraction of exosomes:

[0069] Exosomes were extracted by differential centrifugation. Bone marrow mesenchymal stem cells (BMSCs) were expanded and cultured in a culture dish. When the cell density reached 80%, the original culture medium containing serum was discarded, washed twice with PBS, then added with DMEM, and starved for 48 hours. Collect DMEM containing exosomes, centrifuge at low temperature for three times to remove macromolecular substances, centrifuge for the first time at 4°C, 300g, centrifuge for 10min, absorb the supernatant, discard the sediment, the purpose is to remove substances such as cells and dead cells; Centrifuge for the second time at 4°C, 2000g for 15min, discard the precipitate, and take the supernatant. The purpose of this centrifugation is to remove large cell debris and other substances; The supernatant, the purpose of this centrifu...

Embodiment 2

[0075] Example 2 A preparation method of loaded norcantharidin exosomes and its property characterization

[0076] (1) Extraction of exosomes:

[0077] Exosomes were extracted by ultracentrifugation. Bone marrow mesenchymal stem cells (BMSCs) were expanded and cultured in a culture dish. When the cell density reached 90%, the original culture medium containing serum was discarded, washed with PBS three times, added DMEM, and starved for 45 hours. Collect the DMEM containing exosomes, centrifuge three times at low temperature to remove macromolecular substances, centrifuge for the first time at 6°C, 400g, centrifuge for 5min, absorb the supernatant, discard the sediment, the purpose is to remove cells and dead cells and other substances; Centrifuge for the second time at 6°C, 1000g, for 20min, discard the precipitate, and take the supernatant. The purpose of this centrifugation is to remove large cell debris and other substances; The supernatant, the purpose of this centrifug...

Embodiment 3

[0082] Example 3 A preparation method of loaded norcantharidin exosomes and its property characterization

[0083] (1) Extraction of exosomes:

[0084]Exosomes were extracted by ultracentrifugation. Bone marrow mesenchymal stem cells (BMSCs) were expanded and cultured in a culture dish. When the cell density reached 80%, the original culture medium containing serum was discarded, washed with PBS three times, added DMEM, and starved for 50 h. Collect the DMEM containing exosomes, centrifuge three times at low temperature to remove macromolecular substances, centrifuge for the first time at 2°C, 200g, centrifuge for 20min, absorb the supernatant, discard the sediment, the purpose is to remove substances such as cells and dead cells; Centrifuge for the second time at 2°C, 3000g, for 10min, discard the precipitate, and take the supernatant. The purpose of this centrifugation is to remove large cell debris and other substances; The supernatant, the purpose of this centrifugation ...

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Abstract

The invention belongs to the field of new dosage forms of pharmaceutical tumor drugs, and particularly relates to a preparation method and application of norcantharidin-loaded exosomes. The method specifically comprises the following steps of (1), extracting mesenchymal stem cell exosomes by adopting an ultracentrifugation method; and (2), wrapping norcantharidin in the exosomes through an electroporation method to obtain the drug-loaded exosomes. The prepared drug-loaded exosomes can improve the liver targeting property of the drug and enhance the drug effect of norcantharidin.

Description

technical field [0001] The invention belongs to the field of new formulations of pharmaceutical tumor drugs, and in particular relates to a norcantharidin-loaded exosome and a preparation method thereof. Background technique [0002] With the rapid development of nanotechnology, researchers have been committed to the development of new carriers that can significantly improve drug targeting, such as liposomes, nanoparticles, nanospheres, etc., these carriers take advantage of their small particle size (<1μm ), which can be taken up by organs rich in reticuloendothelial system, such as liver, spleen, bone marrow, etc., thus forming passive targeting. Or it can be structurally modified or transformed so that it has a special affinity for certain tissues and organs, so as to achieve active targeting effects. However, after a large number of animal and clinical trials, it has been found that nanocarriers also have many disadvantages: nanocarriers, as exogenous substances, may...

Claims

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Application Information

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IPC IPC(8): A61K9/50A61K31/365A61K47/46A61K35/00
CPCA61K31/365A61K9/5068A61P35/00
Inventor 王婴王岩梁乐谊赵玲郭密密
Owner GUANGDONG PHARMA UNIV
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